Field of the Invention
The present invention relates to compositions, methods and kits for the species-specific identification of Pseudomonas aeruginosa, which may be present either alone or as a component, large or small, of a homogeneous or heterogeneous mixture of nucleic acids in a sample taken for testing, e.g., for diagnostic testing, for screening of blood products, for microbiological detection in bioprocesses, food, water, industrial or environmental samples, and for other purposes.
Description of the Related Art
The detection and/or quantitation of specific nucleic acid sequences is an important technique for identifying and classifying microorganisms, diagnosing infectious diseases, detecting and characterizing genetic abnormalities, identifying genetic changes associated with cancer, studying genetic susceptibility to disease, measuring response to various types of treatment, and the like. Such procedures are also useful in detecting and quantifying microorganisms in foodstuffs, water, industrial and environmental samples, seed stocks, and other types of material where the presence of specific microorganisms may need to be monitored.
Nucleic acid amplification assays are well suited for the detection of microorganisms in the context of clinical laboratory testing, bioprocess monitoring, or any other setting in which the detection of a specific microorganisms in a particular sample type is desired, by offering high sensitivity and rapid time-to-result relative to conventional microbiological techniques. In addition, amplification methods can be used in the detection of the vast number of microorganisms that are difficult or impossible to culture on synthetic media. Nevertheless, there are limitations associated with these approaches, many stemming from the high level of sensitivity of nucleic acid amplification methods and the resulting amplification of unintended side-products.
Pseudomonas aeruginosa is a gram-negative bacteria that infects humans and can be particularly difficult to treat. In addition, this organism is one of several known contaminant organisms in many biopharmaceutical process streams. Therefore, sensitive and rapid methods for detecting and monitoring the presence of Pseudomonas aeruginosa and other related organisms are continually being sought. While the rapid and accurate detection and/or quantitation of Pseudomonas aeruginosa is highly desirable, it has been difficult to achieve in practice using conventional reagents and techniques. For example, laboratory culture techniques involve incubating samples for 24-48 hours to allow the organisms to multiply to macroscopically detectable levels. Subculture techniques and metabolic assays are then required to distinguish Pseudomonas aeruginosa from related pseudomonads and other enteric bacteria and may require an additional 24-48 hours.
Accordingly, there remains a need in the art for a rapid and robust detection system that can specifically and selectively identify Pseudomonas aeruginosa in a sample of interest. As describe further herein, the present invention meets these needs and offers other related advantages.